Process of extracting suprarenalin.



UNITED STATES g rsn'r OFFICE.

ALBERT G. MANNS AND FREDERICK C. KOCH, OF CHICAGO, ILLINOIS, ASSIGNORS TO ARMOUR & COMPANY, OF CHICAGO, ILLINOIS, A COR- PORATION OF ILLINOIS.

PROCESS OF ExTnAcTme. SUPRARENALIN.

Specification of Letters Patent.

Patented Aug. 21, 1906.

Applicati'in filed December 23.1904. serial No. 238,076. a

county of Cook and State of Illinois, have in vented a certain new and useful Process of Extracting Suprarenalin from the Sn rarenal Glands of Vertebrate Animals, of w ich the following is a specification. r

This invention relates to a new and useful process of producing an extract of suprarenal glands in a physiological active and dry pulverulent or crystalline form.

Such extracts have heretofore been made and are variously known as suprarenalin, epinephrin, or adrenalin, &c, and the product of the present recess is in a general sense similar to the lEefore-mentioned extracts, although it possesses certain characteristics of its own distinguishing it from other analogous products.

The object of the present invention is to provide a simpleand practical process where- )y the extract may be economically secu ed and a relatively lar e quantity obtained from a iven weight of g ands.

n carrying out our invention we first steep the finely-bashed glands in a dilute aqueous solution of acetic acid for a considerable period of time at a low temperature-say from 100 to 112 Fahrenheit. We next coagulate the proteid and matters contained in the fluid extract by heating the same to boiling.- We then filter and concentrate the filtrate in vacuum to a thick syrup; next dilute the syru b coholic solution to a thick syrup in vacuum; then precipitate the active principle from this syrup by adding an aqueous solution of alkyl amins, such as monomethylaminordimethylamin or triinethylamin until the solution becomes distinctly alkaline and gives off the the addition of four to five volumes of a co 01 (preferably wood alcohol) andagain filter; then again concentrate this al finely and then steep the same in a five-percent. solution of acetic acid and water, usin about two ounds of solvent for each pound of bashed g ands and steeping for about two hours at a temperature of about 100 to 112 Fahrenheit.

The liquid extract is drawn off and the glands again steeped for a like period in a like solution for about the same lengthof time, I

and this operation is repeated until the glands have been subjected to four steepings, the li uid extract being in each case retained. The dilute a ueous acid extracts are then heated to the oiling-point and thereafter filtered rapid] 'In this connection is to be noted that all work connected with the steeping and filtering must be done as rapidly as possible and with as'little exposure to the air as practicable. The extracts should not be allowed to come in contact with iron vessels at least for any length of time.

We next evaporate the filtrates in avacuum to such degree of concentration that for every "one hundred pounds of glands there remains from two to three pounds of liquid. To the thick syrup obtatined by this concentration we now radually add, while constantly stirring, t e alcoholic washings from a previous batch (derived as explained below) and as much more ninety-five per cent. wood-alcohol as will equal four and one-half times the volume of the concentrated syrup to which it is added. The solution is then set aside infilled closely sealed or corked bottles and allowed to stand for a considerable period-say twelve hours. After thus standmg the solution is filtered as rapidly as possible and thereafter eva orated to separate out the alcohol. The residue left on the filterpa or is transferred to a crock or other suitab e nonoxidizing vessel and thoroughly mixed with recovered wood-alcohol, (that which has been reviously used for this purpose is preferab e',) which alcohol must test slightly acid toward litmus aper and if found alkaline must be acidified by the addition of a few drops of acetic acid. It will be found desirable to use about one quart of alcohol for the residue derived from each one hundred pounds of su rarenal glands. This mixtureis filtered and the filtrate set aside, the residue being a ain thoroughl mixed with fresh recovere alcohol and t e alterwhich is set in ice-water, so as to cool the nate mixing and filtering repeated until the l finally sucked dry. The moist precipitate residue has been thus washed five times. I from the. filter is now transferred to a Hirsch The filtrates from these five washings are funnel provided with hardened filterpaper, then mixed to be added to the thick syrup, 1 and in effecting this transfer the precipitate 18 as described in connection with the first pre- 1 washed fromt e Buchner filter by the use of a ci itation ith wood-alcohol, small stream of ninety-fiveper cent. wood- The combined alcoholic filtrates are evapoi ,alcohol. This wood-alcohol 1S drawn through rated in a vacuumpan, preferably provided l the Hirsch filterby suction and the precipitate with a condenser and receiver to recover the I washed three times more in the same manner alcohol. The evaporation is continued untila l with ninety-fit e per cent' wood-alcohol, thick syrup remains, which is just sufficiently using, say, one-half pint of wood-alcohol with fluid to pour readily. WVe now add to the each washing. (The alcohol used in these syrup a small uantity of water-say about 1 washings is preserved for future use after it fifty per cent. by volunieand evaporate has YedlstlllBd-l T0 almost y again to the sam n i t ncy the syrup had precipitate one-half pound of light naphtha before the addition of the water. We then is added, allowed to soak through the preagain add water'to such extent as to dilute the lpltate, and Sucked through the filter until syrup sufliciently to permit free filtration by the precipitate is dry. This 0 eration of suction and then filter the liquid, preferably 1 adding small quantities of nap tha 18 reby the use of a large Buchner porcelain funpeated five times, the naphtha being saved nel-say of ten inches diameter-using sueand rcdistilled. FQllOWlIlg the last washing tion to facilitate the filtering. Small quanwlth naphtha air is drawn through the mass tities 0f watermay be used to wash the filuntil the precipitate assumes the form of a ter, taking care to keep the volume of the dry powder. liquid as small as possible. This filtrate liq- The mother liquid, which is obtained by uid is now transferred to a preci itation-'ar, filtration after the solution has been treated (preferably a large wide-mout ed bott e,) with the alkyl amin and allowed to stand, is tested to determine whether or not'it still contains. unprecipitated suprarenalin. This is done b adding a small amount of alk-y] amin whifi; maintained at a low temperature and stirred continuously. If upon the addition of such alkyl ainin further precipitate is formed, the mother liquid is set aside in the chill-room and allowed to standfrom twelve to twenty-four hours and then again decanted and filtered, as in the first instance.

filtrate rapidly, the liquid being stirred thoroughly to facilitate even and ra id cooling. After the li uid has been thoroughly cooled we add in sma amounts at a time a ten-per-cent. solution of alkylamin, stirring well after each addition. The amin is added in this way repeatedly until the liquid gives off a distinct odor of the amin. After a vigorous stirring the recipitation of the extract will take place. In t is connection it is to be noted that the We claim as our inventionprecipitation jar is kept in ice-water during 1. An improvement in the art of extracting the precipitation. The jar is now covered the active princi 1c of suprarenal glands, and set aside in a chill-room and allowed to which consists in st steeping the glands in stand for another twelve or fifteen hours. an aqueous solution at a temperature below The next ste is to filter off and wash the that atwhich albumen coagulates,next coagu precipitate, an in this connection it is imlating the proteid materials by bringing the portant that the operation of filtering, washsolution to a coagulating temperature, and mg, and drying be performed as rapidly and separating the coagulated substances from with as litt e exposure to the air as practicathe solution, then condensing the solution by ble. In filtering the clear supernatant liquid evaporation to a syrupy consistence, then diis first decanted off into a Buchner filter and luting the solution with alcohol and separatfiltered ra idly by suction. To the suping the resultant precipitant, then again conrarenin sed iment left in the precipitation-'ar centrating the fi trate by evaporation to a is added a small quantity of quite cold issyrupy consistence, then precipitating the tilled water and the solution thoroughly active principle by the addition thereto of mixed. After the decanted li uid has pracalk l amin and finally separating the pre tically all passed throu h the uchner filter cipitate from the liquid. this filtrate is set asi e, and the mixture 2. Animprovement in the art of extracting of water and precipitate is then placed in a filter and filtered by suction, as before. When nearly all of the liquid has passed throu h, but the filter-paper is still covered with uid, another quart or so of the cold distilled water is added and allowed to run through, and this addition of small uantities of water is repeated until it has een performed about five times, when the filter is powdered form, W ich consists in first steeping the glands in a slightly-acidified aqueous solution at a temperature below that at which albumen coagulates, next coagulating the pro teid materials by bringing the solution to a coagulating temperature, and separating the coagulated substances from the solution, next con ensing the solution by evaporation to a the active principle of suprarenal glands in saaaao ulated substances from the solution. then condensing the solution by evaporation in vacuum to a syrup consistency,'next diluting the solution wit alcohol and filtering the resultant precipitant therefrom. then again concentrating the filtrate by eva oration in vacuum to a syrupy consistency, t en precipitating the active principle by the addition thereto of alkyl amin. and finally recoverin the precipitate by filtering it from the liquid and drying the same.

A. An improvement in the art of extracting the active principle of suprarenal glands in powdered form, which consists in repeatedly steeping the glands in a slightly-acidified aqueous solution at a temperature below that at which albumen coa ulates, next coa lat ing, the proteid materials by bringing t e solution derived from the several steepings to a coagulating temperature for a brief period of time, next separating the coagulated substances from the solution, then condensing the solution b evaporation in vacuum, next diluting the so ution with alcohol and filtering out the resultant recipitant, then again concentrating the trate by evaporation to a syrupy consistency then chilling the solution and precipitating the active principle by the addition t ereto of alkylamin, then filtering out the resulting precipitate from the liquid then washing the precipitate with water an a volatile liquid in succession and finally drying the powder.

5. An improvement in the art of extracting the active princi le of su rarenal glands, which consists in st steeping the lands in an aqueous solution and se arating t e liquid from the residue, adding a cohol to the solution and filtering the resultant precipitant therefrom, concentrating the solution by evaporation in vacuum, and then preci itating the principle by the addition to t e filtrate of alkyl amin, and finally separating the precipitant from the solution.

6. The improvement in the art of extracting the active principle of suprarenal glands which consists in precipitating said principle from a concentrated aqueous solution of the glands by adding theretoalkyl amin.

7. The improvement in the art of extract ing the active principle of suprarenal glands, which consists in first steeping the glands in an aqueous solution at a temperature below that at which albumen coagulates, next coagulating the proteid materials by bringing the solution to a coagulating temperature, then separating the coagulated substances from the solution, then condensing the solution by evaporation to a syrupy consistency, then adding alcohol to precipitate inert organic substances and separating the resultant precipitant, then again concentrating the filtrate by evaporation to a syrupy consistency, then precipitating the active principle by the addition to the solution of one of the methylamins, and finally separating the precipitate from the liquid.

.ALBERT G. MANNS.

FREDERICK C. KOCH.

Witnesses:

ALBERT H. GRAVES, FREDERICK C. Goopwm. 

